Anion-exchange (AEX) columns were prepared by on-column polymerization of acrylates and methacrylates containing tertiary amino or quaternary ammonium groups on monolithic silica in a fused silica capillary modified with anchor groups. The columns provided a plate height (H) of less than 10μm at optimum linear velocity (u) with keeping their high permeability (K=9–12×10 −14 m 2 ). Among seven kinds of AEX columns, a monolithic silica column modified with poly(2-hydroxy-3-(4-methylpiperazin-1-yl)propyl methacrylates) (HMPMA) showed larger retentions and better selectivities for nucleotides and inorganic anions than the others. The HMPMA column of 410mm length produced 42000–55000 theoretical plates (N) at a linear velocity of 0.97mm/s with a backpressure of 3.8MPa. The same column could be employed for a fast separation of inorganic anions in 1.8min at a linear velocity of 5.3mm/s with a backpressure of 20MPa. In terms of van Deemter plot and separation impedance, the HMPMA column showed higher performance than a conventional particle-packed AEX column. The HMPMA column showed good recovery of a protein, trypsin inhibitor, and it was applied to the separation of proteins and tryptic digest of bovine serum albumin (BSA) in a gradient elution, to provide better separation compared to a conventional particle-packed AEX column.